ID | 65997 |
著者 |
Ren, Jianchao
Department of Chemistry, Okayama University
Danchana, Kaewta
Department of Chemistry, Okayama University
Sasaki, Keiko
Department of Earth Resources Engineering, Graduate School of Engineering, Kyushu University
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抄録 | Laccase is a lignin-degrading enzyme that is expected to move industrial applications to a greener form of biotechnology. Here, we used 2,2'-azinobis(3-ethylbenzthiazolin-6-sulfonic acid) (ABTS) as a mediator and N-benzoyl leucomethylene blue (BLMB) as a substrate to develop a fluorometric assay that we used to measure laccase activity in mushroom extracts. We then compared this novel approach to conventional absorption spectrophotometry. With this novel approach, laccase oxidizes ABTS to produce ABTS radicals that show an absorption maximum at 415 nm. The ABTS radicals oxidize BLMB to generate fluorescent methylene blue that is measured by fluorometry while absorption spectrophotometry directly measures the absorbance of the ABTS radicals at 415 nm. Under the optimal conditions, the fluorometric assay showed a linear calibration curve with limits of detection and quantification of 1.0 × 10-2 mg mL-1 and 3.2 × 10-2 mg mL-1, respectively, and those values are 1.4-fold lower than the results using conventional absorption spectrophotometry to measure ABTS radicals. Laccase activity of extracts from species of mushrooms that include eryngii and shiitake were successfully determined via both fluorometry and absorption spectrophotometry. The eryngii extract showed the highest level of activity, which was followed by the shiitake extract, but laccase activity was not observed in the shimeji extract.
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キーワード | Laccase
Mushroom
Fluorometry
2,2'-Azinobis(3-ethylbenzthiazolin-6-sulfonic acid)
N-Benzoyl leucomethylene blue
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備考 | © 2023 Elsevier Inc. This manuscript version is made available under the CC-BY-NC-ND 4.0 license https://creativecommons.org/licenses/by-nc-nd/4.0/
This fulltext file will be available in Aug. 2024.
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発行日 | 2023-10
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出版物タイトル |
Journal of Food Composition and Analysis
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巻 | 123巻
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出版者 | Elsevier BV
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開始ページ | 105627
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ISSN | 0889-1575
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資料タイプ |
学術雑誌論文
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言語 |
英語
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OAI-PMH Set |
岡山大学
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著作権者 | © 2023 Elsevier Inc.
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論文のバージョン | author
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DOI | |
Web of Science KeyUT | |
関連URL | isVersionOf https://doi.org/10.1016/j.jfca.2023.105627
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ライセンス | https://creativecommons.org/licenses/by-nc-nd/4.0/
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